Recovery of DNA barcodes from blackfly museum specimens (Diptera: Simuliidae) using primer sets that target a variety of sequence lengths
In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies (Diptera: Simuliidae). We analysed 271 pinned specimens representing two genera and at least 36 species. Due to the age of our material, we targeted overlapping DNA...
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| Veröffentlicht in: | Molecular ecology resources 2014-05, Vol.14 (3), p.508-518 |
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| container_title | Molecular ecology resources |
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| creator | Hernández‐Triana, L. M Prosser, S. W Rodríguez‐Perez, M. A Chaverri, L. G Hebert, P. D. N Ryan Gregory, T |
| description | In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies (Diptera: Simuliidae). We analysed 271 pinned specimens representing two genera and at least 36 species. Due to the age of our material, we targeted overlapping DNA fragments ranging in size from 94 to 407 bp. We were able to recover valid sequences from 215 specimens, of which 18% had 500‐ to 658‐bp barcodes, 36% had 201‐ to 499‐bp barcodes and 46% had 65‐ to 200‐bp barcodes. Our study demonstrates the importance of choosing suitable primers when dealing with older specimens and shows that even very short sequences can be diagnostically informative provided that an appropriate gene region is used. Our study also highlights the lack of knowledge surrounding blackfly taxonomy, and we briefly discuss the need for further phylogenetic studies in this socioeconomically important family of insects. |
| doi_str_mv | 10.1111/1755-0998.12208 |
| format | Article |
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M ; Prosser, S. W ; Rodríguez‐Perez, M. A ; Chaverri, L. G ; Hebert, P. D. N ; Ryan Gregory, T</creator><creatorcontrib>Hernández‐Triana, L. M ; Prosser, S. W ; Rodríguez‐Perez, M. A ; Chaverri, L. G ; Hebert, P. D. N ; Ryan Gregory, T</creatorcontrib><description>In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies (Diptera: Simuliidae). We analysed 271 pinned specimens representing two genera and at least 36 species. Due to the age of our material, we targeted overlapping DNA fragments ranging in size from 94 to 407 bp. We were able to recover valid sequences from 215 specimens, of which 18% had 500‐ to 658‐bp barcodes, 36% had 201‐ to 499‐bp barcodes and 46% had 65‐ to 200‐bp barcodes. Our study demonstrates the importance of choosing suitable primers when dealing with older specimens and shows that even very short sequences can be diagnostically informative provided that an appropriate gene region is used. 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M</creatorcontrib><creatorcontrib>Prosser, S. W</creatorcontrib><creatorcontrib>Rodríguez‐Perez, M. A</creatorcontrib><creatorcontrib>Chaverri, L. G</creatorcontrib><creatorcontrib>Hebert, P. D. N</creatorcontrib><creatorcontrib>Ryan Gregory, T</creatorcontrib><title>Recovery of DNA barcodes from blackfly museum specimens (Diptera: Simuliidae) using primer sets that target a variety of sequence lengths</title><title>Molecular ecology resources</title><addtitle>Mol Ecol Resour</addtitle><description>In this study, we evaluated the efficacy of various primers for the purpose of DNA barcoding old, pinned museum specimens of blackflies (Diptera: Simuliidae). We analysed 271 pinned specimens representing two genera and at least 36 species. Due to the age of our material, we targeted overlapping DNA fragments ranging in size from 94 to 407 bp. We were able to recover valid sequences from 215 specimens, of which 18% had 500‐ to 658‐bp barcodes, 36% had 201‐ to 499‐bp barcodes and 46% had 65‐ to 200‐bp barcodes. Our study demonstrates the importance of choosing suitable primers when dealing with older specimens and shows that even very short sequences can be diagnostically informative provided that an appropriate gene region is used. Our study also highlights the lack of knowledge surrounding blackfly taxonomy, and we briefly discuss the need for further phylogenetic studies in this socioeconomically important family of insects.</description><subject>Animals</subject><subject>blackflies</subject><subject>COI</subject><subject>Diptera</subject><subject>DNA</subject><subject>DNA barcoding</subject><subject>DNA Barcoding, Taxonomic - methods</subject><subject>DNA primers</subject><subject>DNA Primers - genetics</subject><subject>Electron Transport Complex IV - genetics</subject><subject>genes</subject><subject>Insect Proteins - genetics</subject><subject>insects</subject><subject>Molecular Sequence Data</subject><subject>museum specimens</subject><subject>Museums</subject><subject>Phylogeny</subject><subject>Simuliidae</subject><subject>Simuliidae - classification</subject><subject>Simuliidae - enzymology</subject><subject>Simuliidae - genetics</subject><subject>taxonomy</subject><issn>1755-098X</issn><issn>1755-0998</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU9v1DAQxSMEoqVw5gaWuJRDWk8cO3FvpVsK0rIgSkVvlu1Mtm7zZ7Gdwn4EvjVJt90DF_BlLOv33oznJclLoAcwnkMoOE-plOUBZBktHyW725fH23t5uZM8C-GaUkFlkT9NdrI8kzIHuZv8_oq2v0W_Jn1NZotjYrS3fYWB1L5viWm0vambNWmHgENLwgqta7ELZH_mVhG9PiLnrh0a5yqNb8kQXLckKz8yngSMgcQrHUnUfomRaHKrvcN41yzgjwE7i6TBbhmvwvPkSa2bgC_u615y8f7028mHdP757OPJ8Ty1uRBlWgkjOfBSGl5ya6uCV5k2pgBRWWORZ6yqcwEls3XBjAZkUKOtKKegQRjG9pL9je_K9-MEIarWBYtNozvsh6CAZ3nOGBTiP1AQTEoQMKJv_kKv-8F340cmKi-AUcZH6nBDWd-H4LFW06q0XyugagpUTZGpKT51F-ioeHXvO5gWqy3_kOAI8A3w0zW4_pef-nS6eDBONzoXIv7a6rS_UaJgBVffF2fqslzM8i_zTL0b-dcbvta90kvvgro4zyhwSmk5DcP-AEB4w0U</recordid><startdate>201405</startdate><enddate>201405</enddate><creator>Hernández‐Triana, L. 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We were able to recover valid sequences from 215 specimens, of which 18% had 500‐ to 658‐bp barcodes, 36% had 201‐ to 499‐bp barcodes and 46% had 65‐ to 200‐bp barcodes. Our study demonstrates the importance of choosing suitable primers when dealing with older specimens and shows that even very short sequences can be diagnostically informative provided that an appropriate gene region is used. Our study also highlights the lack of knowledge surrounding blackfly taxonomy, and we briefly discuss the need for further phylogenetic studies in this socioeconomically important family of insects.</abstract><cop>England</cop><pub>Blackwell Publishing Ltd</pub><pmid>24299419</pmid><doi>10.1111/1755-0998.12208</doi><tpages>11</tpages></addata></record> |
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| ispartof | Molecular ecology resources, 2014-05, Vol.14 (3), p.508-518 |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | Animals blackflies COI Diptera DNA DNA barcoding DNA Barcoding, Taxonomic - methods DNA primers DNA Primers - genetics Electron Transport Complex IV - genetics genes Insect Proteins - genetics insects Molecular Sequence Data museum specimens Museums Phylogeny Simuliidae Simuliidae - classification Simuliidae - enzymology Simuliidae - genetics taxonomy |
| title | Recovery of DNA barcodes from blackfly museum specimens (Diptera: Simuliidae) using primer sets that target a variety of sequence lengths |
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